This study investigated the effect of vitamin B₆ deficiency on antioxidant enzyme activities and lipid profile in rats with exercise-induced oxidative stress. Forty eight rats were fed either a vitamin B₆ deficient diet (B6-) or a control diet (control) for 4 weeks and then subdivided into 3 groups: pre-exercise (PreE); post-exercise (PostE); recess after exercise (recessE). Compared to those of control group, plasma catalase and hepatic cytosol superoxide dismutase (SOD, EC 1.15.1.1) activities of B6- group were lower regardless of exercise. The ratio of reduced glutathione/oxidized glutathione (GSH/GSSG) of B6- group was lower in PreE and there was no difference between PostE and recessE. The level of malondialdehyde (MDA) of B6- was significantly higher in PreE and PostE. High-density lipoprotein-cholesterol (HDL-C) level of B6- group was lower regardless of exercise. Atherosclerotic index of B6- group was higher in PreE and there was no difference between PostE and recessE. It is suggested that a reduction in antioxidative status caused by vitamin B₆ deficiency may be aggravated under exercise-induced oxidative stress.